Characterization of a novel phage depolymerase specific to Escherichia coli O157:H7 and biofilm control on abiotic surfaces

Do-Won Park; Jong-Hyun Park

doi:10.1007/s12275-021-1413-0

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Characterization of a novel phage depolymerase specific to Escherichia coli O157:H7 and biofilm control on abiotic surfaces: Do-Won Park , Jong-Hyun Park; Journal of Microbiology 2021;59(11):1002-1009.
DOI: https://doi.org/10.1007/s12275-021-1413-0
Published online: October 6, 2021

Department of Food Science and Biotechnology, Gachon University, Seongnam 13120, Republic of Korea

Corresponding author: Jong-Hyun Park , Tel: +82-31-750-5523,

Received: 29 July 2021 • Revised: 24 August 2021 • Accepted: 6 September 2021

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Abstract

The increasing prevalence of foodborne diseases caused by Escherichia coli O157:H7 as well as its ability to form biofilms poses major threats to public health worldwide. With increasing concerns about the limitations of current disinfectant treatments, phage-derived depolymerases may be used as promising biocontrol agents. Therefore, in this study, the characterization, purification, and application of a novel phage depolymerase, Dpo10, specifically targeting the lipopolysaccharides of E. coli O157, was performed. Dpo10, with a molecular mass of 98 kDa, was predicted to possess pectate lyase activity via genome analysis and considered to act as a receptor- binding protein of the phage. We confirmed that the purified Dpo10 showed O-polysaccharide degrading activity only for the E. coli O157 strains by observing its opaque halo. Dpo10 maintained stable enzymatic activities across a wide range of temperature conditions under 55°C and mild basic pH. Notably, Dpo10 did not inhibit bacterial growth but significantly increased the complement-mediated serum lysis of E. coli O157 by degrading its O-polysaccharides. Moreover, Dpo10 inhibited the biofilm formation against E. coli O157 on abiotic polystyrene by 8-fold and stainless steel by 2.56 log CFU/coupon. This inhibition was visually confirmed via fieldemission scanning electron microscopy. Therefore, the novel depolymerase from E. coli siphophage exhibits specific binding and lytic activities on the lipopolysaccharide of E. coli O157 and may be used as a promising anti-biofilm agent against the E. coli O157:H7 strain.

Keywords: Escherichia coli O157:H7, bacteriophage, depolymerase, lipopolysaccharide, biofilm, food application

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Characterization of a novel phage depolymerase specific to Escherichia coli O157:H7 and biofilm control on abiotic surfaces

J. Microbiol. 2021;59(11):1002-1009. Published online October 6, 2021

DOI: https://doi.org/10.1007/s12275-021-1413-0

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